Human CCL2/MCP1 ELISA Kit EZ-SetTM (DIY Antibody Pairs)

Monocyte chemoattractant protein-1 (MCP-1), a member of the chemokine (chemotactic cytokine) family, is a potent monocyte agonist that is upregulated by oxidized lipids. MCP-1 is also known as CCL2, SCYA2, MCAF. MCAF is a member of family of factors involved in immune and inflammatory responses. The amino acid sequence deduced from the nucleotide sequence reveals the primary structure of the MCAF precursor to be composed of a putative signal peptide sequence of 23 amino acid residues and a mature MCAF sequence of 76 amino acid residues. MCP-1 plays a unique and crucial role in the initiation of atherosclerosis and may provide a new therapeutic target in this disorder. Human MCP-1 is a 8.7KDa non-glycoprotein, consisting of 99 amino acids in precursor form and 76 amino acids in mature form.

C-C motif chemokine 2; HC11; Monocyte chemoattractant protein 1; Monocyte chemotactic and activating factor; MCAF; Monocyte chemotactic protein 1; MCP-1; Monocyte secretory protein JE; Small-inducible cytokine A2; CCL2; MCP1, SCYA2

Intra-Assay Precision Inter-Assay Precision Sample 1 2 3 1 2 3 n 16 16 16 24 24 24 Mean (pg/ml) 31 159 477 30 148 481 Standard deviation 1.55 8.74 32.91 1.98 9.76 37.51 CV (%) 5% 5.5% 6.9% 6.6% 6.6% 7.8% Intra-Assay Precision Inter-Assay Precision Sample 1 2 3 1 2 3 n 16 16 16 24 24 24 Mean (pg/ml) 31 159 477 30 148 481 Standard deviation 1.55 8.74 32.91 1.98 9.76 37.51 CV (%) 5% 5.5% 6.9% 6.6% 6.6% 7.8% Intra-Assay Precision Inter-Assay Precision Intra-Assay Precision Inter-Assay Precision Sample 1 2 3 1 2 3 Sample 1 2 3 1 2 3 n 16 16 16 24 24 24 n 16 16 16 24 24 24 Mean (pg/ml) 31 159 477 30 148 481 Mean (pg/ml) 31 159 477 30 148 481 Standard deviation 1.55 8.74 32.91 1.98 9.76 37.51 Standard deviation 1.55 8.74 32.91 1.98 9.76 37.51 CV (%) 5% 5.5% 6.9% 6.6% 6.6% 7.8% CV (%) 5% 5.5% 6.9% 6.6% 6.6% 7.8%

Chemotactic factor that attracts monocytes and basophils but not neutrophils or eosinophils. Augments monocyte anti-tumor activity. Has been implicated in the pathogenesis of diseases characterized by monocytic infiltrates, like psoriasis, rheumatoid arthritis or atherosclerosis. May be involved in the recruitment of monocytes into the arterial wall during the disease process of atherosclerosis.

Catalog number Description Quantity Storage of opened/reconstituted material EZ0441-CA Rabbit anti- human CCL2 polyclonal antibody (Capture Antibody) 500 μl, 0.4 mg/mL (recommended dilution 1:100) Store undiluted at 4°C for 1 month or at -20°C for 3 months provided this is within the expiration date of the kit. EZ0441-DA Biotinylated goat anti- human CCL2 polyclonal antibody (Detection Antibody) 500 μl, 5 μg/mL (recommended dilution 1:100) AR1103 Avidin-Biotin-Peroxidase Complex (ABC) 500 μl (recommended dilution 1:100) EK0441-ST Lyophilized recombinant human CCL2 standard 10 ng/tubex3 Discard the standard stock solution after 12 hours at 4°C. May be stored at -20°C for 48 hours provided this is within the expiration date of the kit. Catalog number Description Quantity Storage of opened/reconstituted material EZ0441-CA Rabbit anti- human CCL2 polyclonal antibody (Capture Antibody) 500 μl, 0.4 mg/mL (recommended dilution 1:100) Store undiluted at 4°C for 1 month or at -20°C for 3 months provided this is within the expiration date of the kit. EZ0441-DA Biotinylated goat anti- human CCL2 polyclonal antibody (Detection Antibody) 500 μl, 5 μg/mL (recommended dilution 1:100) AR1103 Avidin-Biotin-Peroxidase Complex (ABC) 500 μl (recommended dilution 1:100) EK0441-ST Lyophilized recombinant human CCL2 standard 10 ng/tubex3 Discard the standard stock solution after 12 hours at 4°C. May be stored at -20°C for 48 hours provided this is within the expiration date of the kit. Catalog number Description Quantity Storage of opened/reconstituted material Catalog number Description Quantity Storage of opened/reconstituted material EZ0441-CA Rabbit anti- human CCL2 polyclonal antibody (Capture Antibody) 500 μl, 0.4 mg/mL (recommended dilution 1:100) Store undiluted at 4°C for 1 month or at -20°C for 3 months provided this is within the expiration date of the kit. EZ0441-CA Rabbit anti- human CCL2 polyclonal antibody (Capture Antibody) 500 μl, 0.4 mg/mL (recommended dilution 1:100) Store undiluted at 4°C for 1 month or at -20°C for 3 months provided this is within the expiration date of the kit. EZ0441-DA Biotinylated goat anti- human CCL2 polyclonal antibody (Detection Antibody) 500 μl, 5 μg/mL (recommended dilution 1:100) EZ0441-DA Biotinylated goat anti- human CCL2 polyclonal antibody (Detection Antibody) 500 μl, 5 μg/mL (recommended dilution 1:100) AR1103 Avidin-Biotin-Peroxidase Complex (ABC) 500 μl (recommended dilution 1:100) AR1103 Avidin-Biotin-Peroxidase Complex (ABC) 500 μl (recommended dilution 1:100) EK0441-ST Lyophilized recombinant human CCL2 standard 10 ng/tubex3 Discard the standard stock solution after 12 hours at 4°C. May be stored at -20°C for 48 hours provided this is within the expiration date of the kit. EK0441-ST Lyophilized recombinant human CCL2 standard 10 ng/tubex3 Discard the standard stock solution after 12 hours at 4°C. May be stored at -20°C for 48 hours provided this is within the expiration date of the kit.

Store at 4°C for 6 months, at -20°C for 12 months. Avoid multiple freeze-thaw cycles (Ships with gel ice, can store for up to 3 days in room temperature. Freeze upon receiving.)

Concentration (pg/ml) 0 15.6 31.2 62.5 125 250 500 1000 O.D. 0.008 0.1 0.206 0.426 0.826 1.271 1.762 2.087 Concentration (pg/ml) 0 15.6 31.2 62.5 125 250 500 1000 O.D. 0.008 0.1 0.206 0.426 0.826 1.271 1.762 2.087 Concentration (pg/ml) 0 15.6 31.2 62.5 125 250 500 1000 Concentration (pg/ml) 0 15.6 31.2 62.5 125 250 500 1000 O.D. 0.008 0.1 0.206 0.426 0.826 1.271 1.762 2.087 O.D. 0.008 0.1 0.206 0.426 0.826 1.271 1.762 2.087

1. Microplate reader in standard size. 2. Automated plate washer. 3. Incubator. 4. Adjustable pipettes and pipette tips. Multichannel pipettes are recommended in the condition of large amount of samples in the detection. 5. Clean tubes and Eppendorf tubes. 6. 96 well microplate (Cat# AR1100) 7. Plate Sealers. 8. Capture Antibody Diluent: PBS. 9. Reagent Diluent: 1% BSA in PBS, pH 7.2-7.4, 0.2 μm filtered. 10. Substrate Solution: Tetramethylbenzidine (Cat# AR1104) 11. Stop Solution: 2 N H2SO4 (Cat# AR1105) 12. Wash Buffer (PBS and PBS-T) . Recipes: PBS: 8g NaCl, 0.2g KCl, 1.15g Na2HPO4, 0.2g KH2PO4, adjust the total volume to 1 L with distilled water, pH 7.2-7.4, 0.2 μm filtered. PBS-T: 0.1% Tween® 20 in PBS, pH 7.2-7.4. *Item 6 – 12 are included in the EZ Set Accessory Kit (EZA001)